title: Ether lipid biosynthesis: isolation and molecular characterization of human dihydroxyacetonephosphate acyltransferase.
abstract: In this paper we describe isolation and molecular characterization of human dihydroxyacetonephosphate acyltransferase (DAP-AT). The enzyme was extracted from rabbit Harderian gland peroxisomes and isolated as a trimeric complex by sucrose density gradient centrifugation. From peptide sequences matching EST-clones were obtained which allowed cloning and sequencing of the cDNA from a human cDNA library. The nucleotide-derived amino acid sequence revealed a protein consisting of 680 amino acid residues of molecular mass 77187 containing a C-terminal type 1 peroxisomal targeting signal. Monospecific antibodies raised against this polypeptide efficiently immunoprecipitated DAP-AT activity from solubilized peroxisomal preparations, thus demonstrating that the cloned cDNA codes for DAP-AT.
authors.affiliation: Biochemie-Zentrum, Universität Heidelberg, Germany.
authors.forename: T P
authors.initials: TP
authors.lastname: Thai
authors.forename: H
authors.initials: H
authors.lastname: Heid
authors.forename: H R
authors.initials: HR
authors.lastname: Rackwitz
authors.forename: A
authors.initials: A
authors.lastname: Hunziker
authors.forename: K
authors.initials: K
authors.lastname: Gorgas
authors.forename: W W
authors.initials: WW
authors.lastname: Just
mesh_terms: D000217:Acyltransferases / Q000032:analysis / Q000737:chemistry* / Q000302:isolation & purification / Q000378:metabolism; D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D001921:Brain / Q000201:enzymology; D002499:Centrifugation, Density Gradient; D003001:Cloning, Molecular; D004591:Electrophoresis, Polyacrylamide Gel; D006243:Harderian Gland / Q000201:enzymology; D006801:Humans; D008830:Microbodies / Q000201:enzymology*; D016253:Microscopy, Immunoelectron; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000737:chemistry / Q000276:immunology / Q000302:isolation & purification; D010742:Phospholipid Ethers / Q000378:metabolism; D011233:Precipitin Tests; D021382:Protein Sorting Signals / Q000737:chemistry; D011817:Rabbits; D017422:Sequence Analysis, DNA
chemical_list: D010446:Peptide Fragments; D010742:Phospholipid Ethers; D021382:Protein Sorting Signals; D000217:Acyltransferases; C020051:glycerone-phosphate O-acyltransferase